The primary objectives of this Program Project are to establish the functional roles of the endocannabinoid system in normal physiological processes and in abnormal or disease states and to determine the extent to which it serves as a mediator in the effects of exogenous cannabinoids. It is our premise that understanding the endogenous cannabinoid system will allow us to address public health issues regarding drugs of abuse as well as co-morbidities that include cognitive disorders, compulsive behavior, pain, motor dysfunction and numerous peripheral disorders. Incredible progress in the past decade has firmly established the existence and homeostatic importance of the endocannabinoid system and its key components (receptor subtypes, endogenous ligands, synthetic and metabolic pathways, signaling pathways, etc.) As with most biological systems, as knowledge accumulates so does the level of complexity. We have assembled a team of chemists (Mechoulam, Razdan and Mahadevan), biochemists and molecular biologists (Cravatt and Di Marzo), and pharmacologists (Martin, Wiley, Lichtman, Pertwee, and Ross) who will address fundamental questions related to distinguishing between the physiological functions of anandamide (AEA) and 2- arachidonoylglycerol (2-AG), investigating whether other endocannabinoids exist, and determining the relationship between the roles of endocannabinoids in neural processes (e.g., pain, reward, neuroprotection) and peripheral processes (e.g., bone formation, inflammation). Our approach involves synthesis of putative endocannabinoids, stable and potent analogs, metabolic and synthetic enzyme inhibitors and CB1 receptor allosteric modulators. Furthermore, we will manipulate enzymes that are putatively responsible for AEA and 2-AG synthesis and degradation as well as establish the resulting lipid profiles using liquid chromatography, ion trap, time-of-flight mass spectrometry to identify new lipid entities associated with the endocannabinoid system. Dr. Cravatt will systematically delete existing as well as proposed synthetic and metabolic enzymes for AEA and 2-AG in mice. These genetically modified animals, along with the synthetic probes and newly discovered endocannabinoids, will be used to further elucidate the function of the endocannabinoid system in neuropathic and inflammatory pain, neurotrauma, reward, and dependence-related events. Ultimately, the knowledge gained from this basic research will yield novel therapeutic targets that can be exploited with the pharmacological agents developed here. PROGRAM CHARACTERISTICS